The Germination Requirements of Spores of Clostridium botulinum Type E

Abstract

The requirements for, and conditions influencing, the germination of spores of Clostridium botulinum type E were studied in chemically defined media. Spores of this organism germinated rapidly and completely in various combinations of L-alanine and one of such carbon sources as sugars, lactate, and ribosides in the presence of bicarbonate. The most effective compounds tested were glucose, L-and D-lactate, and inosine. The formation of lactic acid was found to occur during germination of spores in a combination of L-alanine and glucose. Furthermore, the germination in this medium was strongly inhibited by fluoride or iodoacetate. The germination of spores was also initiated with high concentrations (100 mm or more) of L-alanine or certain other amino acids alone in the presence of bicarbonate at pH 9.0. The spores germinating in L-alanine alone and those in L-alanine plus other compounds were compared as to the minimal concentration of L-alanine required, pH optimum, temperature optimum for heat-activation, bicarbonate requirement, and effects of various inhibitors. Significant differences among these characteristics led to a conclusion that different metabolic pathways, operating selectively each germinative compound used, might be involved in the germination of spores of the organism. The mode of action of combination in stimulating germination by the different germinative compounds has been discussed.

Diversity of the Germination Apparatus in Clostridium botulinum Groups I, II, III, and IV

“…Spores of C. botulinum Group II also respond to a variety of amino acids ( Table 1 ) (Ando and Iida, 1970; Ando, 1971; Plowman and Peck, 2002). In contrast to C. botulinum Group I, L -lactate is considered essential for amino acid induced germination in C. botulinum Group II (Plowman and Peck, 2002).…”

“…In contrast to C. botulinum Group I, L -lactate is considered essential for amino acid induced germination in C. botulinum Group II (Plowman and Peck, 2002). The germination responses of C. botulinum Group II is dependent on strain, pH, temperature, and buffer (Ando and Iida, 1970; Ando, 1971; Plowman and Peck, 2002; Peck, 2009). …”

Use of a novel method to characterize the response of spores of non-proteolytic Clostridium botulinum types B, E and F to a wide range of germinants and conditions

“…L ‐alanine, L ‐cysteine, L ‐serine, L ‐threonine) induced germination of spores of non‐proteolytic Cl. botulinum type E (strain Iwanai) at pH 9·0 in the presence of NaHCO 3 (Ando 1971). At neutral pH, a situation more relevant to food, germination by a single amino acid was moderate, but multi‐component mixtures triggered extensive germination (Ando and Iida 1970).…”

“…At neutral pH, a situation more relevant to food, germination by a single amino acid was moderate, but multi‐component mixtures triggered extensive germination (Ando and Iida 1970). Subsequent work confirmed that while L ‐alanine (25 mmol l −1 )/NaHCO 3 (60 mmol l −1 ) in phosphate buffer (40 mmol l −1 , pH 6·8) had no effect, further addition to the mixture of glucose, galactose, maltose, glucose‐6‐phosphate, fructose‐1,6‐diphosphate, L ‐lactate, D ‐lactate, DL ‐lactate, guanosine or inosine at 2–20 mmol l −1 induced germination of spores of strain Iwanai (Ando 1971). A number of alpha‐hydroxy acids were also effective (Ando 1974).…”

“…Limited information indicates that spores of non‐proteolytic Cl. botulinum type E strain Iwanai respond to a combination of nutrient germinants (Ando and Iida 1970; Ando 1971, 1974). Lysozyme brings about enzymatic germination of spores of non‐proteolytic Cl.…”